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Information on EC 2.7.1.119 - hygromycin-B 7''-O-kinase and Organism(s) Streptomyces hygroscopicus and UniProt Accession P09979

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IUBMB Comments
Phosphorylates the antibiotics hygromycin B, 1-N-hygromycin B and destomycin, but not hygromycin B2, at the 7''-hydroxy group in the destomic acid ring.
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Streptomyces hygroscopicus
UNIPROT: P09979
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The taxonomic range for the selected organisms is: Streptomyces hygroscopicus
The enzyme appears in selected viruses and cellular organisms
Synonyms
hygromycin phosphotransferase, hygromycin b phosphotransferase, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hygromycin B phosphotransferase
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HPH
-
-
-
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HPT
-
-
-
-
hygromycin B phosphotransferase
-
-
-
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hygromycin phosphotransferase
-
-
-
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phosphotransferase, hygromycin B
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-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
phospho group transfer
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-
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SYSTEMATIC NAME
IUBMB Comments
ATP:hygromycin-B 7''-O-phosphotransferase
Phosphorylates the antibiotics hygromycin B, 1-N-hygromycin B and destomycin, but not hygromycin B2, at the 7''-hydroxy group in the destomic acid ring.
CAS REGISTRY NUMBER
COMMENTARY hide
88361-67-5
-
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
ATP + hygromycin B
ADP + 7''-O-phosphohygromycin B
show the reaction diagram
-
-
-
?
ATP + hygromycin B
ADP + 7''-O-phosphohygromycin B
show the reaction diagram
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
ATP + hygromycin B
ADP + 7''-O-phosphohygromycin B
show the reaction diagram
-
-
-
?
ATP + hygromycin B
ADP + 7''-O-phosphohygromycin B
show the reaction diagram
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
hygromycin B
substrate inhibition
hygromycin B
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above 0.01 mM, substrate inhibition
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0423 - 0.0822
ATP
0.0167 - 0.03
hygromycin B
0.0364 - 0.076
ATP
0.00056 - 0.0015
hygromycin B
additional information
additional information
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-
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2.5 - 5.6
ATP
6.4 - 8
hygromycin B
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
59.1 - 68.1
ATP
267 - 383
hygromycin B
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.61 - 1.27
hygromycin B
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.6 - 8.5
-
pH 6.6: about 40% of maximal activity, pH 8.5: about 65% of maximal activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
gene hyg
UniProt
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION?
KHYB_STRHY
332
0
37054
Swiss-Prot
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MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
41000
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1 * 41000, SDS-PAGE
42000
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gel filtration
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
x * 37980, wild-type enzyme, sequence calculation, x * 38348, enzyme mutant HYG10, sequence calculation
monomer
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1 * 41000, SDS-PAGE
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
L79F/S83L/G189R/E237K/S252P/D285N/L290M
random mutagenesis, mutant HYG10 contains seven substitutions and a duplication, it is thermostabilized compared to the wild-type enzyme, three-dimensional structure analysis, overview. Substitutions G189R, E237K, and D285N are located at the protein surface. The mutant shows altered kinetics at 45°C compared to the wild-type enzyme
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
35
T1/2 of wild-type enzyme HYG
48 - 74
T1/2 of mutant enzyme HYG10, Tmax is 74°C
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
glycerol and bovine serum albumin stabilize
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20°C, in 50% v/v glycerol and 0.1% bovine serum albumin, stable for up to 6 months
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PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant His6-tagged enzyme fom Escherichia coli strain BL21(DE3) by nickel affinity and anion exchange chromatography, and gel filtration
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
gene hyg, subcloning in Escherichia coli strain JM109, recombinant expression of His6-tagged enzyme in Escherichia coli strain BL21(DE3), recombinant enzyme expression from plasmid pT8S-P31 in Thermus thermophilus strain B27 TH104 providing hygromycin B resistance up to 60°C, while recombinant expression of a mutant variant HYG10, containing seven substitutions and a duplication, provides HgB resistance up to 74°C
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Zalacain, M.; Pardo, J.M.; Jimenez, A.
Purification and characterization of a hygromycin B phosphotransferase from Streptomyces hygroscopicus
Eur. J. Biochem.
162
419-422
1987
Streptomyces hygroscopicus
Manually annotated by BRENDA team
Pardo, J.M.; Malpartida, F.; Rico, M.; Jimenez, A.
Biochemical basis of resistance to hygromycin B in Streptomyces hygroscopicus - the producing organism
J. Gen. Microbiol.
131
1289-1298
1985
Streptomyces hygroscopicus
Manually annotated by BRENDA team
Sugimoto, N.; Takakura, Y.; Shiraki, K.; Honda, S.; Takaya, N.; Hoshino, T.; Nakamura, A.
Directed evolution for thermostabilization of a hygromycin B phosphotransferase from Streptomyces hygroscopicus
Biosci. Biotechnol. Biochem.
77
2234-2241
2013
Streptomyces hygroscopicus (P09979), Streptomyces hygroscopicus
Manually annotated by BRENDA team